Organic oxidation products and process of preparing the same



' Patented 193 1935 I oxoamc OXIDATION rnonuc'rs rnocsss on rasranmo m SAME mm W. um, Chicago, and Herbert out Ron-' 1 UNITED smras PATENT" "obi-leg I nor, Des Plalnes, Ill., assign era, by means as'- signmentajowJsRfShort Milling Company,

. Ghicago, 111., a corporation of Illinois.

No Drawing. Application December 27, 1932,

. Serial No. 649,110

29 Claims. (01. 260 16) tion of organic compounds. to any extent up to a' '\practical or theoretical limit is accomplished by contacting oxidizable organic-compositions and gas containing oxygen in.- the presence of.

- enzymic material con g; enzymes which 'ei'' feet the catalytic oxidation of such compositions.

In the process of thepresent invention, oxide It is found that .the longer the period of treat ment of certain compoundsfwith air, oxygen or ozone, the greater is the degree of oxidation.

The degree of oxidation oian oxidizableorganic I material is readily determinedby quantitative" methods and may be expressed in terms of mole- .cules (M) '01 hydroge peroxideper 1000 grams of organic material or grams of hydrogen per-'- oxide per one hundred grams of organic matter. The so-called 'M-values are given below in thefirst mentioned terms. 1 I

Besides increasing the M-value of an organic composition by lengthening the time of treatment with oxygen, this value may also-be increased by the'addition of enzymic material, and by increas ing the amounts of cozy-mic material in contact with the, organic materialftreatedl lit-is now shown that certain enzymic materials under given .conditions have a strong catalyzing effect which 40 cannot be obtained under similar conditions by the use of air or oxygen alone. The catalyzing,

effect is likewise not obtainable by mere oommingling oiairor oxygen with oil in the presence of water. An increase in concentration of enzymic -'material results in a corresponding increase in M-values-so long as the mixture of enzymic material and organic mattertreated' permits of the most intensive incorporation of oxygen or air. It

- is readily seen that in order to obtain maximum 5o oxidation 1 with air, for instance, as the oxygen' source; the system to be treated must at all times be supplied with as much air as possible, and the air must be brought in contact with the largest Various materials may be employed for supplying enzymes, and more particularly. oxidizing .en'-.

'zymesincluding peroxidase, oxidase, catala'se', and

perhydridase, to be used as c'atalysts-inthe present process. Soya bean meal has b en ioundto be 5 an inexpensive and a convenien catalyst in the enzyme oxidation of organic materials. Other leguminous materials, including peas (iorinstance yellow peas) and beans (for instance navy beans) I or mixtures of leguminous materials may be used. 10 I L Extracts of leguminous materials may be employed to advantage in place of whole seed jflour or meal. Such extracts may be employed either prior 3 I to or after clarification. In the case of soya beans, either the soya bean milk or the casein-free milk 15 or extractmay be used; As to-the. flour. or other source herein mentioned, itshould be, noted thatsuch flour or other source'isnot ordinary commercial material, but especially prepared so as to retain its enzyme value and therefore be 20 factive" within the-meaning or this specification. a

;Exmcrrox or En zxsuc'Acms able ior 'use in the present process may be pre- 25 pared as follows: Enzyme-active whole soya beans are washedand s'oakedin an excess'(2 or 3 times the quantity oi. beans) of cold wateror' preferably ice water for about six to sixteen hours 0r long'eny The soaked beans are ground fine to so permit of the mostcomplete leaching, out oi the enzymes by water without clogging. the pores of a filter used for separating the solid residue from the liquid; About 2000 parts by weight of ground,

soaked beans are immersed in aboutf6500 parts as by .weightoi cold water and the liquid is pressed out by suitable means after a iew minutes of soaking,'thereby producing a millw extract. About 20 parts by weight of calcium chloride are dissolved in water and added'to the soya bean milk, while 40 stirring. After a thorough mixing, the precipitate V formed is allowed to settle for several hours The I clarified upematant liquid is" decanted and -fil erecl.

The proportion of water used in the ex- 45 traction process .maybe varied, but care should be taken that all the soyacasein precipitated by -calcium chlorideis removed,- and the consistency oi the'casein-rree extract does not interiere with itsprepar'ation and-use. 'It is advantageous to obtain as high a-concentration' of oxidizing" 811- ,zymes as possible in an extract. As stated above the unclarifled extract'ot son I possible surface of the organic material' v treated' beans, or. soya bean milk-mayI be usedin my} to within a given unit oi.time.

I, procesain which case thetre'atnientsvitli- I extracts containing enzymes suitchloride is omitted. However, the use of the milk. makes it difficult to separatefinely divided matter,

added with the milk, from a treated organic composition to obtain a relatively pure. enzyme oxiwhen using soyabean milki firhe stepsgepessary to prepare casein -free-extracts become un ieces :h sary when employing beans and peas. Thus, thea whole extractionprocedure greatly simplified and the preparation of more concentrated extracts isnade possible/ A navy bean extract i prepared as follows: 500 parts by weight "of na beans are soaked for about sixteen hours in pproximately 2000 parts.

by weightot cold water. The soaked beans are ground and the groundmaterial is immersed in the soaking water, The extract is separated from centrifuging,

Fatty acid compounds including the acids themselves or the corresponding esters are readily oxidizable by the present process. Animal or vegetable oils or fats are suitable sources for 80 such compounds. For instance, cocoanut oil, cottonseed oil, corn oil, poppyseed oil, butter, lard, hydrogenated fats or oils, and fatty acids or esters derived from the oils and fats mentioned may be treated. Olic acid, for example commercial 85 grades, is oxidizable by ,the present process.

Inthe description of the present process, the termsoil and fatmaybeused interchangeably. Wherever the term aeration is used, it may be construed to mean the exposure of preferably all portions of the mixto air, pure oxygen or ozone, oxygen or ozone diluted. with air, nitrogen, or other inert gas. Chemicals 'such as hydrogen the pulp by decantation orby filtration or by v with a suspension of approximately 40 parts by weight of active'spya flour in 200 parts by weight peroxide may be employed as a source of oxygen.

Oxygen may be liberated under the'influence of catalase, for instance, which is usually contained in certain sources of oxidizing enxymes. Instead of agitating the mix by means of a mechanical stirrer, agitation may be accomplished by the oxy' gen-carrying gas.

Aeration may be performed in an open or in a closed vessel, or inother words under atmospheric or super-atmosphericpressure. Higher pressures are advantageous especially when higher concentrations of owgen are employed.

, Oxidation in the presence of enzymes is favorably affected by higher temperatures than room.

temperatures, as'for instance up to substantially 50 C. e

/ As indicated above, contact is made between oxidizable organic compositions and gas contain;

: ing free oxygen in the presence of enzymic material. It is understood that any medium that -makes this contact possible may be used and that such a medium acts asa carrier or dispersing agent for the reacting substances without impair- 1 of cotton seed oil, partly hydrogenated (having' an original M-value of 0.0095), and a suspension of parts by weight of jactivef soya flour in about 360 parts by weight of water, is thoroughly aerated by beating vigorously forabout 240min'-'--= utes at'about to about (48.8? to 60 C.) The fatty material thus treate is then separated from the other constituents of the mix by centrifuging or by extraction with suitable solvents.

A sample of the product obtained by this process 10 employing centrifugal 'separation showecLaneM- .A.

value of 0.1670. A sample of the product oxidized -lnthe presence of enzymes isolated by extraction withethyt or petrolic ether, and heating the extract to dryness until the residue reached atemperatureof'about 220 F. (104.4 C.) showedan M-value of 0.1503. Thisdifferenee in the M-values is due to the increased temperature employed in drying the extracted product. The diiference is quite. small however, and indicates that the V enzyme oxidized product is rather'stableI 0M Example II About 138 partsby weight of corn oil are mixedof water. To this mixture are added about 3 parts by weight of 40% hydrogen peroxide.- The whole mix, is intensively aerated for about forty-five minutes. The protein matter (casein) in the're- 3o sulting mix is then separated by coagulation, or decreasing or completely offsetting. its swelling capacity. A few parts by weight of 40 f rmaldehyde are added and the mix is heated to b'oiling,

" whereupon two layers form on standing.- The top 35 layer is removed, freed of the bulk of the water therein by centrifuging, and salt worked in, which causes exudation of the oxidized oil. The'oil is decanted and is recovered as far as possible by centrifuging and pressing the residue. 4 40 A sample of oil thus obtained showed an M- value of 0.1654, indicating that hydrogen peroxide facilitates and accelerates the enzymicoxidation.

After 45 minutes of aeration, a higher M-value is obtained than after 240 minutes in Example I. 45 a In Example If, it is found thatformaldehyde may be used as a coagulant for proteins without apparent harm to the oxidizing power of the treated oil.

The amount of soya flour used varies preferably 5 between about 16 and 30% of the amount of oil. Expressed in terms of percent of the total mix, the amount of soya flour used in Examples I and II.

is-practically the same (12% and 10% respective ly). The proportion of enzymic material'in the 55 mix may obviously bev'aried within very wide limits, depending to a large extent on the efiicie'ncy of thedevices employed for agitation or aeration, and the degree of oxidation desired or obtainable under given conditions. The proportions of other 5 materials used may also be widely varied Example III About 1380 parts by weight of corn oil, preferably refined, and about '3000'parts by weight of 65 I soya 'beanextract prepared in the -manner described above, are thoroughly aerated for about four and.one-half hours, a small amountof'pure The resulting material is .7

separation into layers takes place. The top layer is removed, freed of water by centrifuging, or decantation, and aerated for. about four hours at about 120 t0130 I". in the presenceof about 5 and a moderate supply of pure oxygen. The enzyme oxidizedfattymaterial may be separated,

' from the-treated mix as follows: The mix isalface and to form a top layer.

lowed to stand (if necessary after-addinga' little formaldehyde in aqueous solution) inorder to hasten the rising of the oil and casein to the'sur- The top layer, is

4 then separated fromlthe lower layer .and sub? jected to reduced pressure in order to remove" gas-(air or oxygen) which is finely distributedthroughout the material. The gas-free material.

Y is centrifuged to separate water from the oil and protein matter.

protein by treatment with salt and then by de- The oil is separated from the canting or centrifuging and pressing of the res- A sample er oxidized oil thus obtained'showed an M-value of 0.50. The oil may be purified bycentrifugingheating on a water bath and then drying in vacuo. In contrast to the brownish yellow color of the original corn oil, the purified oxidized oil isof a pale lime-yellowish color. A sample of purified oil showed an -M- value of 0.4 4;

"Example IV About EGO/parts by weight of an extract of navybeans prepared in the manner described above, are mixed with about 138 parts by weight of corn oil and the mixture intensively aerated at about 110 to 120 F. for abouttwo hours; Pure oxygen is bubbled through the emulsion during Due to the presence of only small the last hour. amounts of mucilaginous matter in the navy bean extract in contrast to soybean extract, the mix foams only slightly. This behavior greatly facilitates the separation of the treated, oil, which is accomplished byheating the whole mixture to boiling after addition of a very small amount of diluted formaldehyde, then centrifuging, working the upper of two layers obtained thereby with salt, and then pressing the liquid from the resulting mixture, A sample of clear oil thus obtained, showed an M'-value of 0.225,

- Example V About 360 parts by weight of soya oil, 360

parts by weight of water and 150 parts by weightof soya flour are mixed together and thoroughly aerated. Water is added if the mix gets too stiff during aeration. At the end of four hours of aeration, about 400 parts by. weight of water and 30 parts by weight of fresh soya. bean flour, are added to the mix to obtain the,consistency.of

thick cream. The aeration is continued and thevcreamy consistency maintained by adding water from time to time and also by adding fresh soya fiourat intervalsof for instance every two hours. The soya oil is finally clarified and puri fied. At the end of fourteen hours? aeration, a sample ofoil showed an M-value of .5074,

In the above examples, crushed sesame seeds 1 may be added to the mix in addition to soya bean flour or'extract. Thistresults in an increase in the Mvalue, the value being higher than that been flour alone as a source of enzymes. For instance the aeration of a mix comprising 300 four 'hours resulted in an oxidized product having an M-value nearly 7% higher than that obtained A without sesame seeds afterrseven hoursaeration.

In'the preparation of enzyme oxidized prod- I ucts by this process certain characteristics of enzyme oxidized fats or oils in organic solvents.

such as petrolic or, ethyl ether until after the evaporation of the solvent and until a temperature of- 220- F. is reached; only slightly affects their M-value.

As to the use of oxidized. fats in dough bleaching processes, reference is made to our copending idue ina hydraulic press or the like between filter cloth.

application, Serial No. 654,304 filed January 30, As there pointed out, the products of the present invention are readily miscible with shortenlngs. These mixtures 'or the enzyme oxidized enings to be used" for baking purposes. Such shortenings, while thus employed, although they serve to some extent to bleach dough during mixing and .fermentationexert their main bleaching actionduring baking of the dough.

The invention. has been disclosed herein for illustrative purposes in its preferred embodi-- ment, but it is to be understoodthat the scope of- 20 products themselves may be employed as shortfree oxygen in the presence. 'of enzymic material 4 containing enzymes to effect the catalytic oxi-I dation of the organic matter, the said oxidizable organic matter being selected from a group consisting of animaloils and fats, vegetable oils and fats, hydrogenated oils and fats, and fatty acids and esters from said oils and fats. v I

2. A fatty; acid composition oxidized in the presence of enzymic material containing oxidizing of preparing organic oxidation enzymes.

3,..A process products, which process comprises contacting an oxidizable fatty acid composition and gas containing free oxygen in the presence of enzymio materialcontaining enzymes to effect the catalytic oxidation of the composition.

4. A vegetable fatty material oxidized in the presence of enzymic material containing oxidizing enzymes.

'5.'A process of preparing organic oxidation product's, which process comprises contacting vegetable fatty material and gas containing free oxygen in the presence of enzymic material containing enzymes to effect the catalytic oxidation of the fatty;gmaterial. i

6: An animal fatty material treated with gas containing free oxygen in the presence of enzymic material containing oxidizing enzymes.

7. A process of preparing organic oxidation products, which process comprises contacting animal fatty material and gas containing free of the fat or oil oxidized in the presence of soya oxygen in the presence of enzymic material containing enzymes to effectthe catalytic-oxidation of the fatty material.

8. A *process of preparing organic oxidation .ter,-.thesaid oxidizable'organic matter being selected from a group consisting of animal oi1s and fats, vegetable oilsand fats, hydrogenated oils and fats, and fatty acids and esters from said 16. A process of preparing organic oxidation oils and-fats.

9.A process of preparing organic oxidation products, which process comprises contacting oxidizable organic matter andozo'ne in the presence of enzymic material containing enzymes to effect the catalytic oxidation of the organic matter, the said oxidizable organic matter being se lected from a group consisting of animal oils and fats, vegetable oils and fats, hydrogenated oils and fats, and fatty acids and esters from said oilsandfats. I.

10. A process of preparing organic oxidation products, which process comprises contactingoxidizable organic matter and gas containing free oxygen in the presence of vegetable enzyme material containing enzymes to effect the catalytic oxidationof the organic matter, the said oxidize.- ble organic, matter being selected from a group consisting of animal oils and fats, vegetable oils and fats, hydrogenated oils and fats, and fatty acids and esters from said oils' and fats.

11. A process of preparing organic oxidation products, whichprocess comprises contacting oxidizable organic matter and gas containing free 7 from said oils and fats.

.oilsahdfats,

oxygen in the presence of soya bean enzymic material to efiect the catalytic oxidation of the organic matter,- the said oxidizable organic matter being selected from a group consisting of animal oils and fats, vegetable oils and fats, hydrogenated oils and fats, and fatty acids and esters from 3: said ollsand fats. I

' 12. A process of preparing organic om'dation products, which process comprises contacting oxidizable organic matter and gas containing free oxygen in the presence of navy ,hean, enzymic material to effect the catalytic, oxidation of the organic matter, the said oxidizable organic matter ing, selected from a group cgnsisting of animal o and- "fats, vegetableoils and fats, hydro.

genated oils and fats, and-fatty acids and esters from said 'oils and fats.

13. A process. of preparing organic oxidation products, which process comprises contacting oxidizable organic matter and gas containing free owgen'in the presence of pea enzymic material to efiect the catalytic" oxidation of the organic matter, the saidoxidizable organic matter being selected from a group consisting of animal oils and fats, vegetableoils and fats; hydrogenated oils and fats, and fatty acids and esters from said ILA process of pr'parin'g organicioxidation I products, which process comprises contacting ox idimble organic matter and gas containing free omgen inithe presence of'oxidizing enzyme material to eflect the catalytic oxidation oft-he oranic matter the said oxidizable organic matter being selected from a group consisting of animal oils and fats, vegetable oilsand fats, hydrogen-E ated' oils and fats, and ,fattyacids and esters,

15. A process of preparing organic I oxidation products, which'process comprises dispersing oxidimble' organic matter and enzymic materialin a-liquid medium that does not impair the enzymic reactivity required-for the ensuing reaction, the said oxidizable organic matter-being from agroup of animal'oilsand fats, vege and mm acids and esters from said ofls and 15 table oils and ms, hydrogenated oils and fats;

fats, resulting with gas 'ing substances.

' gas containing free oxygento'oxidize the said products, which process comprises dispersing ox containing free oxygen to oxidize the said organic matter, and separating the enzyme oxidized organic matter from the resulting substances.

products, which process comprises dispersing .ox-' idizable organic 'matter and enzymic material in aliquid medium that does not impair the enzymic reactivity required fortheensuing reactiongthe said oxidizable organic matter being selected from a group consisting of animal oils and fats, vege- 10 I table oils and fats, hydrogenated oils and fats; and fatty acids and esters-from said oils and fats;

contacting the resulting dispersion with oxygen to oxidize the said organic. matter, and separating the oxidized organic matter from the result-15 I 'reaction, the said oxidizable organic matter being selected from a group consisting of animal oils and fats, vegetable oils and fats, hydrogen' ated oils and fats, and'fatt'y acids and esters from said oils and fats, contacting the resulting dispersion with gas containing free oxygen to oxidize the said organic matter, and separating the enzyme oxidized organic matter from the resulting 18. A process of preparing organic oxidation products which process comprises dispersing ox idizable organic matter and navy bean enzymic, matter in a liquid medium that does not impair 'the enzymic reactivity required for theensuing reaction, the said oxidizable organic matter being selected from a group consisting of animal oils andfats, vegetable oi1s and fats, hydrogenated oils and fats, and fatty-acids; and esters from Y said oils and fats, contacting the resulting dispersion with gas containing free oxygen to oxidize the said organic matter, and separating the enzyme oxidized organic matter from the result-.

19. A process. of preparing organicpxidation idizable organic matter and-pea enzymic matter in a liquid medium that does notimpair the enzymic reactivity required for the ensuing reaction, the said oxidizable organic matter beingse lepted from a group'consisting of animal oils and fats, vegetable oils and'fats, hydrogenated oils and fats, and fatty acids and esters from said oils and fats, contacting the resulting dispersion with organic matter, and separating the enzymeloxidized organicmatter from the resulting sub-.-

. products, which process comprises dispersing ox- 7 20. A processiof preparing organic oxidation idizable organic matter andoxidizing enzymes with gas containing free oxygen to oxidize.-the J said organic matter, and the enwme oxidized organic matter the resulting sub 21 A process of preparing organic "oxidation in a liquid medium that does not impair theenp products, which process organic an aqueous 11185 dium containing oxidizing enzyme material, the said aqueous medium serving as a carrier for said enzyme material and the said oxidizable organic matter being selected from agroup consisting of animal oils and fats, vegetable oils and fats, hydrogenated oils and fats, and fatty acids and esters from said oils and fats, adding a small proportion of an inorganic peroxide, contacting the resulting emulsion with gas containingfree oxygen to oxidize the fat or fatty oil material, and separating the enzyme oxidized material from the resulting substances.

22. A'process of preparing a bleaching agent, which process comprises dispersing fatty material and enzymic material containing oxidizing enzymes in a liquid medium that does not impair the enzymic reactivity required for the ensuing reaction, aerating the resulting'mixture to oxidize the fatty material in the presence of the enzymic material, and separating the enzyme oxidized fatty material from the mixture.

23. A process of preparing a bleaching agent, which process comprises mixing fatty material and enzymic material containing oxidizing enzymes with water, aerating the resulting mixture to bring about contact of all the particles of the mixture with oxidizing gas to effect thorough oxidation of the fatty material, and separating the enzyme oxidized fatty material from the mixture.

24. A process of preparing a bleaching agent,

- which process comprises mixing a fatty material and soya flour with water and hydrogen peroxide, aerating the resulting mixture to oxidize fatty matter in the said fatty material, and separating the treated fatty material from. the said mixture.

25. A process of preparing a bleaching agent which process comprises mixing a fatty material, soya flour and crushed sesame seed with water, aerating the resulting mixture to oxidize fatty matter in the said fatty material, and separating the treated fatty material from the said mixture.

26. A process of preparing a bleaching agent, which process comprises dispersing fatty material and enzymic material in a. liquid medium that does'not impair the enzymic reactivity required for the ensuing reaction, aerating theresulting mixture until the M-value of the enzyme oxidized fatty material is substantially increased, and separating the oxidized fatty material from-the mixture. V

27. A fattymaterial having bleaching properties and comprising fatty acid compounds ox-. idized in the presence of oxidizing enzymes.

28. A process of preparing a bleaching agent, which process comprises mixing soya oil and soya flour with water, aerating the resulting mixture to oxidize the said soya oil, and sepa-- rating the enzyme oxidized soya oil from the mixture. i 29. A process of preparing a bleaching agent, which process comprises mixing fatty material and enzymic material containing oxidizing enzymes with water, aerating the resulting mixture whilerepeatedly adding fresh portions of said enzymic material to thereby accelerate oxidization of the fatty material, and separating the enzyme oxidized fatty material from the mix- LOUIS W. HAAS. HERBERT OTTO RENNER. 

